8A4E
Room temperature structure of AtPhot2LOV2 in a photostationary equilibrium
Summary for 8A4E
| Entry DOI | 10.2210/pdb8a4e/pdb |
| Descriptor | Phototropin-2, 6-(3-TETRADECANOIC ACID) FLAVINE MONONUCLEOTIDE, FLAVIN MONONUCLEOTIDE, ... (4 entities in total) |
| Functional Keywords | lov domain, plant protein |
| Biological source | Arabidopsis thaliana (thale cress) |
| Total number of polymer chains | 1 |
| Total formula weight | 16146.95 |
| Authors | Engilberge, S.,Caramello, N.,Royant, A. (deposition date: 2022-06-10, release date: 2023-03-29, Last modification date: 2024-10-23) |
| Primary citation | Aumonier, S.,Engilberge, S.,Caramello, N.,von Stetten, D.,Gotthard, G.,Leonard, G.A.,Mueller-Dieckmann, C.,Royant, A. Slow protein dynamics probed by time-resolved oscillation crystallography at room temperature. Iucrj, 9:756-767, 2022 Cited by PubMed Abstract: The development of serial crystallography over the last decade at XFELs and synchrotrons has produced a renaissance in room-temperature macromolecular crystallography (RT-MX), and fostered many technical and methodological breakthroughs designed to study phenomena occurring in proteins on the picosecond-to-second timescale. However, there are components of protein dynamics that occur in much slower regimes, of which the study could readily benefit from state-of-the-art RT-MX. Here, the room-temperature structural study of the relaxation of a reaction intermediate at a synchrotron, exploiting a handful of single crystals, is described. The intermediate in question is formed in microseconds during the photoreaction of the LOV2 domain of phototropin 2 from , which then decays in minutes. This work monitored its relaxation in the dark using a fast-readout EIGER X 4M detector to record several complete oscillation X-ray diffraction datasets, each of 1.2 s total exposure time, at different time points in the relaxation process. Coupled with UV-Vis absorption spectroscopy, this RT-MX approach allowed the authors to follow the relaxation of the photoadduct, a thio-ether covalent bond between the chromophore and a cysteine residue. Unexpectedly, the return of the chromophore to its spectroscopic ground state is followed by medium-scale protein rearrangements that trigger a crystal phase transition and hinder the full recovery of the structural ground state of the protein. In addition to suggesting a hitherto unexpected role of a conserved tryptophan residue in the regulation of the photocycle of LOV2, this work provides a basis for performing routine time-resolved protein crystallography experiments at synchrotrons for phenomena occurring on the second-to-hour timescale. PubMed: 36381146DOI: 10.1107/S2052252522009150 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.96 Å) |
Structure validation
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